3RDE

Crystal structure of the catalytic domain of porcine leukocyte 12-lipoxygenase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	CCD
Collection date	2011-02-03
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.0781
Spacegroup name	P 1 21 1
Unit cell lengths	83.450, 181.540, 91.610
Unit cell angles	90.00, 92.86, 90.00

Refinement procedure

Resolution	48.968 - 1.892
R-factor	0.17323
Rwork	0.172
R-free	0.21447
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2p0m
RMSD bond length	0.026
RMSD bond angle	1.974
Data reduction software	MOSFLM
Data scaling software	SCALA (3.3.16)
Phasing software	PHASER (2.1.4)
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.968	48.968	1.990
High resolution limit [Å]	1.890	5.980	1.890
Rmerge		0.065	0.284
Number of reflections	214779
<I/σ(I)>	9	8.3	2.4
Completeness [%]	99.5	99.3	98.3
Redundancy	3.6	3.8	3.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	291	protein 6 mg/mL incubated with the solid OPP inhibitor for 16 hours, then further concentrated to 11~12 mg/mL in 10 mM Tris HCl, 1 mM TCEP, pH 7.4 buffer. reservoir solution: 0.1 M MES pH 6.5, 5%-10% PEG-20,000, 20% glycerol against 1 mL well solution of 0.1 M MES, 5%-10% PEG-20,000, 20% glycerol, VAPOR DIFFUSION, HANGING DROP, temperature 291K